| In recent years,as one of acute cerebrovascular disease,stroke has been threatening human health and life.The ischemic stroke accounts for about 80%of all cases.This disease has gained prior attention and encouraged researchers to launch a series of related studies due to its high morbidity,disability and mortality.The compatibility of Salvia miltiorrhiza and Dalbergia odorifera performs exhibits strong effects including activating blood circulation and removing blood stasis,relieving pain,and has been applied to treat cardio-cerebrovascular diseases for years in China.In this study,middle cerebral artery occlusion(MCAO)rat model was made to investigate the effects of Salvia miltiorrhiza and Dalbergia odorifera on anti-free radicals,anti-inflammation and metabolic pathways in vivo.The specific research contents are as follows:1.A HPLC method was stablished to determine the content of Tanshinone IIA,Cryptotanshinone,Tanshinone I and salvianolic acid B in Salvia miltiorrhiza,and Danshensu,protocatechuic acid,protocatechuic aldehyde,caffeic acid,lithospermic acid and rosmarinic acid in the injections.Addtionally,the clarity of the injections was checked and the hemocytolysis of the injections was studied.The results showed that the total contents of tanshinone II_A,cryptotanshinone and tanshinone I in Salvia miltiorrhiza was 0.286%(>0.25%)and 3.31%(>3.0%)for salvianolic acid B,which met the requirements of the Pharmacopoeia(2015).The content of protocatechuic aldehyde in Danshen injection and Xiangdan injection were 413.15 g/mL(>200 g/mL)and 414.45 g/mL(>170 g/mL),respectively,which was in accordance with Ministry of Health Drug S tandards;The prepared injection has good clarity and no hemolysis,indicating that the injections conforms to the clinical safety management standard of TCM injection.2.Rats in each group were intravenously administered for 7 days,and then permanent MCAO rat model was made using the intraluminal filament method.The cerebral water content neurological deficit scores and cerebral infarct volume in rats were observed;meanwhile,the contents of lactic acid(LD),lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde(MDA)in brain tissue were determined using biochemical detesting system;the contents of interleukin-1 beta(IL-1β),interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factor alpha(TNF-α)in brain tissue were measured by ELISA assay.The results showed that Salvia miltiorrhiza combined with Dalbergia odorifera could significantly reduce brain moisture content,cerebral infarction volume and improve neurobehavioral disorders in MCAO rats.At the same time,xiangdan injection could significantly increase the contents of LDH,CAT and SOD in the brain,and reduce the contents of LD and MDA and inflammatory factors IL-1β,TNF-αand IL-8.These effects of xiangdan injection were better than that of Danshen and Dalbergia odorifera.3.In this study,rat urine and blood samples were collected as the research objects,metabolomics based on LC–MS was employed to characterize metabolic alterations in rats of different group.The results showed that 56 differential metabolites were found in urines and plasma samples compared with the sham operation group.Salvia miltiorrhiza-Dalbergia odorifera reversed 36 different substances which significantly changed in MCAO rats,and adjusted disturbed metabolic pathways such as amino acid metabolism,fatty acid adjusted disturbedmetabolicpathwaysbeta-oxidation,glycerophospholipidmetabolism,sphingomyelin metabolism and citric acid cycle. |
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